HUMAN ACIDIC RIBOSOMAL PHOSPHOPROTEINS-P0, PHOSPHOPROTEINS-P1, AND PHOSPHOPROTEINS-P2 - ANALYSIS OF CDNA CLONES, INVITRO SYNTHESIS, AND ASSEMBLY
HUMAN ACIDIC RIBOSOMAL PHOSPHOPROTEINS-P0, PHOSPHOPROTEINS-P1, AND PHOSPHOPROTEINS-P2 - ANALYSIS OF CDNA CLONES, INVITRO SYNTHESIS, AND ASSEMBLY
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DOI：
10.1128/mcb.7.11.4065
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发表时间：
1987-11-01
影响因子：
5.3
通讯作者：
STEITZ, JA
中科院分区：
文献类型：
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作者：
RICH, BE;STEITZ, JA
分离并测序了编码三个抗原性相关的人类核糖体磷酸化蛋白P0、P1和P2的cDNA克隆。P1和P2类似于大肠杆菌核糖体蛋白L7/L12， P0可能是L10的类似物。这三种蛋白具有几乎相同的羧基末端17个氨基酸序列（KEESEESD(D/E)DMGFGLFD-COOH），这是它们免疫交叉反应的基础。P1和P2 cdna编码的氨基酸序列与已发表的同源大鼠、卤虾和酿酒酵母蛋白的初级结构高度相似，证实了它们的身份。通过杂交选择mRNA的翻译和产物的免疫沉淀，初步鉴定了P0 cDNA。为了证明编码序列是完整的，我们在体外用噬菌体T7 RNA聚合酶转录P0、P1和P2 cdna，并在体外翻译得到的mrna。合成的P0、P1和P2蛋白在血清学和电泳上与从HeLa细胞中提取的p蛋白相同。这些合成的p蛋白被整合到60S核糖体中，而不是40S核糖体中，并组装成类似于大肠杆菌L7/L12和L10的复合体。
cDNA clones encoding three antigenically related human ribosomal phosphoproteins (P-proteins) P0, P1, and P2 were isolated and sequenced. P1 and P2 are analogous to Escherichia coli ribosomal protein L7/L12, and P0 is likely to be an analog of L10. The three proteins have a nearly identical carboxy-terminal 17-amino-acid sequence (KEESEESD(D/E)DMGFGLFD-COOH) that is the basis of their immunological cross-reactivity. The identities of the P1 and P2 cDNAs were confirmed by the strong similarities of their encoded amino acid sequences to published primary structures of the homologous rat, brine shrimp, and Saccharomyces cerevisiae proteins. The P0 cDNA was initially identified by translation of hybrid-selected mRNA and immunoprecipitation of the products. To demonstrate that the coding sequence are full length, the P0, P1, and P2 cDNAs were transcribed in vitro by bacteriophage T7 RNA polymerase and the resulting mRNAs were translated in vitro. The synthetic P0, P1, and P2 proteins were serologically and electrophoretically identical to P-proteins extracted from HeLa cells. These synthetic P-proteins were incorporated into 60S but not 40S ribosomes and also assembled into a complex similar to that described for E. coli L7/L12 and L10.